Molecular Cloning And Prokaryotic Expression Of Follicles HGT KAP6.1, 7, 8 From Hetian Sheep

Hetian sheep is a local history-honored species with medium-thickness wool, which can be processed into the famous Hetian carpets and included into national animal genetic protection list in 2006. On the basis of certain genes KAPs affecting the growth of follicles in Hetian sheep, the cause of affecting the quality of wool was investigated by using certain molecular techniques to lay foundation on the theoretical basis of genetic protection and cultivation. Hetian sheep as a important donor for the production of Hetian carpets will directly enhance the quality of Hetian carpets and boost sustainable development of local economy.The total RNA was abstracted from the skin of mountain Hetian sheep using Trizol method and then reverse transcripted by reverse transcriptase and HGT genes KAP6.1,KAP7 and KAP8 was determined and amplified by RT-PCR, which was then inserted into pMD-18T vector. The right recombinant plasmid sequence was corroborated after double digestion of restriction enzyme and PCR determination. The comparison between detected sequence by DNAMAN and sequence from GeneBank showed the follicle genes KAP6.1,KAP7 and KAP8 identified with those from GeneBank.The acquired HGT genes KAP6.1,KAP7 and KAP8 were cloned into prokaryotic expression vector pET28a(+) to assembly targeted prokaryotic expression vector. The acquired prokaryotic expression vector was transformed into Escherichia strain BL21(DE3) and then inducing-cultivated by IPTG. The SDS-PAGE test specific expression band lied in the location about 10KD.